Characterization of a Novel Esterase Belonging to Family V from Marinobacter flavimaris

Lipolytic enzymes have attracted enormous attentions because of their ability in ester hydrolysis,ester synthesis,transesterification and other biochemical reactions.Bacteria are important sources of lipolytic enzymes applied in industry.Here,a novel lipolytic enzyme encoded by esterase gene est1347 was identified in Marinobacter flavimaris WLL162,and was purified and characterized.The lipolytic enzyme Est1347 consisted of 312 amino acid residues and a 21-amino-acids N-terminal signal peptide with a predicted molecular weight of 34.2 kDa.It belongs to family V of bacterial lipolytic enzymes based on the amino acid sequence homology analysis.Est1347 is a mesophilic and alkali-resistant enzyme with the highest activity at 45℃and pH 8.5;it is stable at temperatures below 50℃and pH 7.5–11.0.Est1347 showed a preference for middle-length chain substrate p-NPC10 and a wide range of other substrates.The Km,Vmax,Kcat and Kcat/Km values of Est1347 for p-NPC10 in pH 8.5 at 45℃were 0.9411 mmol L^(−1),1285μmol min^(−1)mg^(−1),698.91 s^(−1)and 743.65 s^(−1)(mmol L^(−1))^(−1),respectively.It is also tolerant to the metal ions,organic solvents and detergents.In conclusion,the esterase Est1347 laid a foundation for further study of bacterial lipolytic enzyme family V.

Thiosulfate oxidation and autotrophy potential by marine prevalent heterotrophic bacteria of genus Marinobacter

The genus Marinobacter is very broadly distributed in global environments and is considered as aerobic heterotroph.In this study,six Marinobacter strains were identified with autotrophic thiosulfate oxidation capacity.These strains,namely Marinobacter guineae M3B^(T),Marinobacter aromaticivorans D15-8PT,Marinobacter vulgaris F01^(T),Marinobacter profundi PWS21^(T),Marinobacter denitrificans JB02H27T,and Marinobacter sp.ST-1M(with a 99.93%similarity to the 16S rDNA sequences of Marinobacter salsuginis SD-14B^(T)),were screened out of 32 Marinobacter strains by autotrophic thiosulfate oxidization medium.The population of cells grew in a chemolithotrophic medium,increasing from 105 cells/mL to 10^(7) cells/mL within 5 d.This growth was accompanied by the consumption of thiosulfate 3.59 mmol/L to 9.64 mmol/L and the accumulation of sulfate up to 0.96 mmol/L,and occasionally produced sulfur containing complex particles.Among these Marinobacter strains,it was also found their capability of oxidizing thiosulfate to sulfate in a heterotrophic medium.Notably,M.vulgaris F01^(T)and M.antarcticus ZS2-30^(T)showed highly significant production of sulfate at 9.45 mmol/L and 3.10 mmol/L.Genome annotation indicated that these Marinobacter strains possess a complete Sox cluster for thiosulfate oxidation.Further phylogenetic analysis of the soxB gene revealed that six Marinobacter strains formed a separate lineage within Gammaproteobacteria and close to obligate chemolithoautotroph Thiomicrorhabdus arctica.The results indicated that thiosulfate oxidizing and chemolithoautotrophic potential in Marinobacter genus,which may contribute to the widespread of Marinobacter in the global ocean.

海洋细菌Marinobacter adhaerens HY-3分离鉴定及对中肋骨条藻的化感作用

菌藻化感作用是一种极其复杂的生理、生态学现象.从虾蟹混养池分离得到1株海洋细菌,经16S rRNA序列比对确定为Marinobacter adhaerens HY-3,选择我国常见赤潮优势种中肋骨条藻(Skeletonema costatum)作为对象,研究海洋细菌与其种间化感效应,利用共培养和添加细菌胞外代谢物,以S.costatum生长量及叶绿素a含量为参数,研究细菌对S.costatum生长及光合作用的影响.结果表明,菌液浓度104以上对S.costatum生长产生抑制,培养至第10 d,104、106、108组藻生长量分别是对照组的70%、23%、22%;叶绿素合成量是对照组88%、62%、60%;抑制效应随菌浓度的增加有逐渐增加的趋势.添加细菌胞外代谢物,对S.costatum生长无影响;因此,M.adhaerens HY-3对S.costatum存在一定的化感作用,对其生长及光合作用均产生较大的影响;S.costatum与M.adhaerens HY-3的种间相互作用,是由菌藻细胞的直接接触进行,代谢物不存在化感作用因子.

Corrosion Inhibition of X80 Steel in Simulated Marine Environment with Marinobacter aquaeolei

In the present study, a novel bacterium( Marinobacter aquaeolei) was examined for its corrosion inhibiting behaviour against X80 pipeline steel. Electrochemical results showed that X80 steel immersed in the solution inoculated with M. aquaeolei possessed very high corrosion resistance compared to that of abiotic control. Besides, scanning electron microscopy, confocal laser scanning microscopy and energy-dispersive X-ray spectroscopy were employed to analyse the corrosion product and the biofi lm formed on the metal surface. Fourier-transform infrared spectroscopy was also applied to determine the composition of extracellular polymeric substances(EPS). Above results indicated that the corrosion inhibition effciency observed in biotic medium was very high(91%), proving that M. aquaeolei was an effective inhibitive agent for the corrosion of carbon steel. The inhibition was credited to the formation of bacterial biofi lm and the compact protective layer of the secreted EPS. Thus, this study will introduce a natural, environmentally friendly and cost-effective system for the corrosion control of the carbon steel.

厌氧及好氧条件下盐度对海杆菌NY-9生长及反硝化能力的影响

从盐城市射阳盐场分离筛选出的一株中度嗜盐兼性厌氧海杆菌（Marinobacter）NY-9，该茵在厌氧及好氧条件下均表现出高效的反硝化能力。研究考察了厌氧及好氧条件下盐度对一株海杆菌NY-9生长及反硝化能力的影响。结果表明．菌株该菌株的厌氧及好氧生长盐浓度范围分别为O％～16％及0％。12％。最适生长盐度均为8％，该盐度下培养72h，OD600mm。值分别达到了0.38及1．12。在最适盐度条件下，该菌株均取得了最高的厌氧及好氧反硝化效率．在NO3-起始浓度为100mg／L的反硝化溶液中培养72h，其总氯去除率分别达到了87．6％及62．5％。